At T0 and T90, semen samples were obtained after three days of sexual abstinence. They were collected into sterile containers and held at room temperature for 30 min to liquefy. After liquefaction, all sperm samples were processed through the two-layer density gradient method (Sydney IVF Sperm Gradient, Cook Medical, Sydney, Australia) according to the manufacturer’s instructions. Basic sperm analysis was carried out according to the WHO (World Health Organization) criteria [16 ], including sperm count, concentration, and motility. Sperm score for motility evaluation was expressed as grades a to d, and progressive motility rate was calculated as the percentage of a + b.
Sydney ivf sperm gradient
Manufactured by Cook Medical
Sourced in Australia
The Sydney IVF Sperm Gradient is a lab equipment product used for the separation and purification of sperm cells from semen samples. It is designed to isolate motile and viable sperm cells for use in various assisted reproductive techniques.
Automatically generated - may contain errors
Lab products found in correlation
Sydney ivf blastocyst medium, by Cook Medical (1 mentions)
Sydney ivf gamete buffer, by Cook Medical (1 mentions)
Sydney ivf cleavage medium, by Cook Medical (1 mentions)
Galaxy 48r incubator, by Eppendorf (1 mentions)
Amiloride, by Merck Group (1 mentions)
Ham s f 10, by Merck Group (1 mentions)
Nigericin, by Merck Group (1 mentions)
Kb r7943, by Merck Group (1 mentions)
4 protocols using sydney ivf sperm gradient
1
Sperm Analysis via Density Gradient
2
IVF Semen Preparation and Embryo Transfer Protocol
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Semen samples were prepared for infertility treatments using two-layer density gradient method (Sydney IVF Sperm Gradient, Cook Medical, Sydney, Australia). In ART cycles, controlled ovarian hyperstimulation, oocyte recovery, embryo culture, and morphological evaluation of embryos were performed as described previously (32 (link)). Oocytes were fertilized with cIVF or ICSI. Embryo transfer (ET) was generally performed 72 h after oocyte collection. However, based on of the number of embryos available, the ET could be performed on day 2 (if the patient had only one to two fertilized oocytes) or day 5 (if the patient had at least four good quality cleavage-stage embryos). Surplus blastocysts were cryopreserved.
The serum beta hCG test was performed 14 days after IUI or oocyte recovery. The presence of a gestational sac at the 6rd-7th gestational week was defined as a clinical pregnancy. Miscarriage was defined as a loss of pregnancy after ultrasonographic detection of a gestational sac.
The serum beta hCG test was performed 14 days after IUI or oocyte recovery. The presence of a gestational sac at the 6rd-7th gestational week was defined as a clinical pregnancy. Miscarriage was defined as a loss of pregnancy after ultrasonographic detection of a gestational sac.
3
Comprehensive IVF Oocyte and Embryo Protocol
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After 2 h of incubation, the oocytes were denuded in HEPES-buffered medium (Sydney IVF Gamete medium, Cook Medical) containing 20 IU/ml of Hyaluronidase (Origio, Målov, Denmark). ICSI was performed immediately after denudation. Sperm samples were treated with a two-layer density gradient system (Sydney IVF Sperm Gradient, Cook Medical) or via Swim-up using Sydney IVF Gamete Buffer (Cook Medical). Incubations were performed at 37 °C in a humidified atmosphere of 6 % CO2, 5 % O2 (Galaxy 48R incubator; New Brunswick Scientific). Fertilization was assessed 16–18 h after injection (PN score 1: symmetrical, score 2: non-symmetrical, score 3: abnormal) [1 (link)], and embryos with two pronuclei were individually cultured from day 1 to day 3 into Sydney IVF Cleavage medium (Cook Medical) and from day 3 to day 5–7 in Sydney IVF Blastocyst medium (Cook Medical).
Standard day 2–3 embryo and blastocyst morphological assessment was carried out according to the current consensus system [1 (link)]. Arrested embryos were non-viable embryos in which development arrested for at least 24 h, or in which all the cells degenerated or fragmented.
Standard day 2–3 embryo and blastocyst morphological assessment was carried out according to the current consensus system [1 (link)]. Arrested embryos were non-viable embryos in which development arrested for at least 24 h, or in which all the cells degenerated or fragmented.
4
Sperm Motility Evaluation with Inhibitors
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Motile spermatozoa were selected from 1 ml of liquefied semen by centrifugation (600 g for 10 min) through a discontinuous Sydney IVF Sperm Gradient (Cook Medical, Bloomington, IN, USA) (0.5 ml of 80% and 0.5 ml of 40%). The supernatant was discarded and the pellet was resuspended with 1 ml of Ham's F-10 (Sigma Chemical), at pH 7.8. After another centrifugation (600 g for 5 min), the supernatant was discarded and the pellet was resuspended in 0.5 ml Ham's F-10. The sperm concentration ranged from 3 to 6 millions/ml. The sperm concentration and motility were determined by Computer Assisted Semen Analysis (CASA; Hamilton Thorne, Beverly, MA, USA). The experiments were carried out by incubating 100 ml of motile spermatozoa with 100 ml of Ham's F-10 at 37 8C, and in agreement with the experimental protocol, in the presence and absence of ouabain, amiloride, nigericin, eosin and KB-R7943 (Sigma Chemical). The sperm motility (as percentage of forward progressives from the total sperm cells analysed), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), beat cross frequency (BCF) and amplitude of lateral head displacement (ALH) were determined.
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